The Effect of β-Carotene Against Adriamycin Toxicity on the Embryo Formation

Adriamycin is an anthracycline antibiotic widely used for the treatment of many types of cancer. The cytotoxic effect of Adriamycin occurs by a free radical-mediated mechanism. Thus, to prevent or reduce the toxic effect of Adriamycin, it is possible to use it in combination with antioxidants. The aim of this study was to evaluate a potential effect of β-carotene against Adriamycin-induced toxicity on the embryo formation. Materials and Methods. Pregnant rats were treated with Adriamycin, β-carotene, and their combination during the critical stages of embryogenesis. The first group was control group. Adriamycin was administered on day 9 (group 2a) and day 12 (group 2b) of gestation by a single intraperitoneal injection at a dose of 5 mg/kg. β-Carotene was given at a dosage of 0.6 mg/(kg.day) from day 6 to 10 or from day 9 (group 3a) to 13 (group 3b) of gestation 5 times per os; in the case of their combination, β-carotene was given per os 3 times before Adriamycin injection, one time simultaneously with Adriamycin and one time after its injection (groups 4a and 4b). Animals were euthanized on day 21 of gestation. Embryo resorptions and alive fetuses were counted, weighed, and measured. The embryos of each litter were examined macroscopically after the Buen solution fixation for the embryo defects. In order to render the skeleton visible, the soft tissues were macerated using caustic soda, stained with alizarin red, and cleared with glycerin. Results. Adriamycin induced embryotoxicity; the combination of Adriamycin and β-carotene decreased the number of Adriamycin-induced embryo resorptions about two times. A gavage with Adriamycin alone decreased fetal body weights (P<0.05), while giving it in combination, the fetal body weight was similar to that in the control group. Adriamycin induced the retardation of skeletogenesis and external fetal malformations (microphthalmia, hydrocephaly, anencephaly, and others). After an exposure to β-carotene, external malformations (diaphragmatic hernia) of embryos were found only occasionally. β-Carotene in combination with Adriamycin produced no positive effect on Adriamycin-induced skeletodysgenesis or external malformations. Conclusions. Antioxidant β-carotene in combination with Adriamycin slightly reduced the Adriamycin- induced embryotoxicity, but produced no positive effect on Adriamycin-induced skeletodysgenesis or external malformations

Treatment of Lewis lung carcinoma by photodynamic therapy and glucan from barley

Objective. During the photodynamic treatment, complement system is activated and tumor cells are opsonized with iC3b fragment. β-glucans can enhance cytotoxicity of iC3bopsonized cells due to their specific interaction with complement receptor 3 (CR3; CD11b/CD18) on the surface of the effector cells. In contrast to microorganisms, tumor cells lack β-glucan as a surface component and cannot trigger complement receptor 3-dependent cellular cytotoxicity and initiate tumor-killing activity. This mechanism could be induced in the presence of β-glucans. This study aimed at determining the influence of coadministration of β-glucan from barley on the efficacy of photodynamic tumor therapy (PDT). Material and methods. C57 Bl/6 female mice bearing Lewis lung carcinoma were used throughout the study. Mice were randomized into groups (15 in each group) and exposed to the treatment with intravenous Photofrin injection (dose, 10 mg/kg) and after 24 h following laser illumination, or with oral administration of β-glucan from barley at a dose of 400 μg/mouse per day up to 5 days, or with their combination. Tumor growth dynamics and survival of the treated and untreated mice were monitored. Results. Tumor volume in all treated groups was significantly lower (P<0.001) than that in the control group. The most effective tumor growth suppression (P=0.033) was achieved in mice treated with combination of PDT and β-glucan from barley as compared with PDT alone. The best survival was achieved in the same group, but difference was not significant as compared to the control group (P=0.143) and to PDT alone group (P=0.319). Conclusions. The present study demonstrates that coadministration of β-glucan from barley can enhance efficacy of photodynamic therapy

mTHPC-mediated photodynamic treatment of Lewis lung carcinoma in vitro and in vivo

Background and objective. The ongoing search for the enhancement of efficacy of photodynamic therapy stimulates the interest in molecular mechanisms of the response to the treatment. Looking for the cell line suitable for investigation of cellular response both in vivo and in vitro, we evaluated phototoxicity of m-tetrakis-(3-hydroxyphenyl)-chlorin (mTHPC) on viability of Lewis lung carcinoma (LLC1) cells in vitro, growth of murine transplantable tumor, and mice survival. Material and methods. LLC1 cell culture and male C57BL/6 mice bearing Lewis lung carcinoma were used for the experiments. Photodynamic treatment was mediated by m-tetrakis-(3- hydroxyphenyl)-chlorin as a photosensitizer. Light emitting diode array was used for illumination. The effect of the photodynamic treatment was evaluated by comparison of viability of control and treated cells, growth of tumors, and survival of the control and treated mice. Results. In vitro, a cytotoxic dose inducing a reduction in viability of LLC1 cells by 50% was achieved at 60 mJ/cm2 and approximately 400 ng/mL of the photosensitizer, or 30 mJ/cm2 and 600 ng/mL of mTHPC. Both the concentration of the photosensitizer and duration of light exposure were significant determinants of cytotoxic effect. In vivo, an injection of 0.25 mg/kg of mTHPC to mice bearing Lewis lung tumor and illumination at 120 J/cm2 taking place after 24 h significantly inhibited tumor growth and prolonged mice survival. However, the tumors regained their growth potential after 9 days. Conclusions. Photodynamic treatment mediated by m-tetrakis-(3-hydroxyphenyl)-chlorin had a significant effect on LLC1 cells in vitro and growth of Lewis lung carcinoma in vivo

Accumulation of photosensitizer in rat embryos (a spectroscopic study)

The aim of the present study was to investigate the accumulation of photosensitizer Photofrin II in the different organs as well as the placenta and embryos of pregnant rats and to determine during which stage of embryogenesis the photosensitizer is accumulated the most effectively. Materials and methods. The experiments were carried out on 25 fetuses from 10 Wistar rats (weight 160-240 g). Female rats were mated with male rats in the evening. Vaginal smears were collected from each female rat next morning and were examined by microscope in order to determine the presence of sperm. The day when sperm was detected in the vagina was considered to be pregnancy day 0. Photofrin II (a dose of 5 mg/kg) was administered intravenously to pregnant rats on days 7, 14, 16, 18 and 20 of embryogenesis. Rats were euthanized 24 hours after intravenous injection of Photofrin II and the following organs were taken: brain, spleen, liver, kidneys, lungs, uterus, placenta, and embryos. The accumulation of the photosensitizer was observed in the samples prepared from these parts of body. Fluorescence measurements ex vivo were performed with an S2000-FI fluorescence spectrometer (Ocean Optics Inc., Florida, USA) by exciting the samples with a blue light emitting diode (λ=400 nm). Results. A comparative study of fluorescence spectra on days 7, 14, 16, 18 and 20 of embryogenesis showed that the most intense accumulation of Photofrin II in the embryo was on day 7, while on the other days of embryogenesis the accumulation of Photofrin II increased obviously in the uterus and placenta. Conclusions. The obtained data show that the accumulation of Photofrin II in the embryo depended on the stage of embryogenesis as well as on permeability of the placental barrier. Further photodynamic therapy studies are necessary to determine the total effect of Photofrin II on the embryo